[Effects of thymus tissue transplantation on the survival of mice exposed to a lethal X-Ray dose.]

During the radiation exposure of the body, processes similar to those characteristic of the natural aging of mammals occur. This phenomenon is most often referred to as accelerated aging. By transplanting the thymus tissue, it is possible to achieve a normalization of food and water consumption, a stabilization of weight gain, and a significant increase in the life expectancy of experimental animals, even after their lethal irradiation.

[Upper gastrointestinal duplication complicated by perforation in a child]. / Udvoenie verkhnikh otdelov zheludochno-kishechnogo trakta, oslozhnennoe perforatsiei u rebenka.

Gastrointestinal duplications are rare congenital malformations occurring in embryonic period of development of digestive system. These abnormalities are usually found in infancy or early childhood. Clinical presentation is extremely diverse depending on dimensions, localization and type of duplication. The authors present duplication of antral and pyloric parts of the stomach, the 1st segment of the duodenum and pancreatic tail. Mother with a 6-month-old child turned to the hospital. According to the mother, the child was sick for about 3 days when episodes of periodic anxiety first appeared. Upon admission, abdominal neoplasm was suspected after ultrasound. On the second day after admission, anxiety increased. There was impairment of appetite, and the child rejected food. Abdominal asymmetry in umbilical area was observed. Considering clinical data on intestinal obstruction, emergency transverse right-sided laparotomy was performed. A tubular structure was found resembling intestinal tube was found between the stomach and transverse colon. Surgeon found duplication of antral and pyloric parts of the stomach, the 1st segment of the duodenum and its perforation. During further revision, additional pancreatic tail was diagnosed. En-bloc resection of gastrointestinal duplications was carried out. Postoperative period was uneventful. Enteral feeding was initiated after 5 days, and the patient was transferred to surgical unit. The child was discharged after 12 postoperative days.

PECULIARITIES OF REMODELING OF HEART AND NEUROHUMORAL STATUS IN PATIENTS WITH CHRONIC HEART FAILURE AND CONCOMITANT RENAL DISEASE DEPENDING ON THE VALUE OF THE EJECTION FRACTION OF THE LEFT VENTRICLE.

67 patients with CHF of FC II and concomitant chronic pyelonephritis at remission stage for not less than 6 months. As of the screening moment, intervention group (n=67 patients) and control group (n=22 apparently healthy individuals) were formed. The intervention group was divided into 3 groups: group 1 included 22 CHF patients with preserved LV EF (≥50%) associated with AH and concomitant CRD. Group 2 included 23 CHF patients with decreased heart LV EF (≤40%, but not less than 35%) associated with AH and concomitant CRD. Group 3 was comprised of 22 patients with CHF associated with AH and concomitant CRD, having LV EF 40-49% (so-called "medium" LV EF). In order to evaluate the condition of RAAS regulators, plasma aldosterone level (PA) and plasma renin activity level (PRA) were studied. The ultrasonic parameters were measured in order to determine structural-functional condition of the heart. the study of structural-functional condition of heart, taking into account both systolic and diastolic function of the left ventricle of heart, comparison of parameters in CHF patients associated with AH and CRD with preserved LV EF and "medium" LV EF, PA (by 31.28%) and decrease of PRA parameter (by 14.5%) in the group of CHF patients associated with AH and CRD with "medium" LV EF. Comparison of the groups of CHF patients associated with AH and CRD with decreased LV EF and "medium" LV EF has identified the increase of such parameters as relative wall thickness index (by 2.13%), MMI (by 6.13%), PA (by 22.4%); besides, the increase of Е/е parameter (by 2.7%) was identified in the group of CHF patients with "medium" LV EF.

THE RELATIONSHIP OF THE IMMUNE AND METABOLIC DISORDERS DURING VARIOUS METHODS OF MULTICOMPONENT GENERAL ANESTHESIA IN LAPAROSCOPIC CHOLECYSTECTOMY.

BACKGROUND: In addition to operating injury in the pathogenesis of immunological and metabolic disorders after surgical interventions anesthesia plays an important role. THE AIM: to establish the relationship of the immune and metabolic disorders during various methods ofmulticomponent general anesthesia in conditions of laparoscopic cholecystectomy in patients with cholelithiasis. MATERIALS AND METHODS: Under constant observation there were 68 women admitted to the hospital for surgical treatment of cholelithiasis. Patients were divided into 3 groups depending on multicomponent general anesthesia (halothane, propofol, sevoflurane). We determined the concentration of cytokines (TNFa, IL-la, IL-i/8, IL-4, IL-iRA, IL-2, IFNy), components of the complement system (C,, C3, C4, C, and C, factor H, C,-inhibitor), the activity of neutrophilsperipheral blood, the concentration of the products ofperoxidation, catalase, superoxide dismutase in blood plasma. RESULTS: The level of immune-inflammation and metabolic disorders in patients with cholelithiasis was higher in patients operated with the use of halothane. The use of sevoflurane has had the most positive effect on the studied indices. CONCLUSION: The close correlation between the investigated immune and metabolic parameters on the background of the use of different schemes of multicomponent general anesthesia in patients with cholelithiasis have let to the conclusion that in the conditions of use of sevoflurane has the least place a "tension" immune and oxidative status.

[Clinical case of Jeune syndrome].

The paper reports a clinical case of Jeune syndrome in a baby, in whom the diagnosis was established immediately after his birth. He was also diagnosed as having multiple congenital malformations, such as Dandy-Walker anomaly, hexadactyly of the left hand, cleft palate, choroidal colobomas in both eyes, and atrial septal defect and he also had interstitial nephritis and hepatitis.

Receptor-independent actions of PPAR thiazolidinedione agonists: is mitochondrial function the key?

Agonists of the peroxisome proliferator activated receptor gamma (PPAR(gamma)) are currently used for treatment of type 2 diabetes due to their insulin sensitizing and glucose metabolism stabilizing effects. More recently some of these same agonists were shown to exert anti-inflammatory and anti-proliferative effects as well. Although PPAR(gamma) agonists can operate via receptor-mediated events occurring at the genomic level, thereby causing long lasting changes in gene expression patterns, recent studies demonstrate non-genomic as well as genomic actions, and receptor-dependent as well as receptor-independent effects of the thiazolidinedione (TZD) class of PPAR(gamma) agonists. In this review we will summarize data describing some of these novel, receptor independent actions of TZDs, review evidence that TZDs directly influence mitochondrial function, and attempt to reconcile how changes in mitochondrial function could contribute to other receptor-independent actions of these drugs.

A 27-bp region of the inducible nitric oxide synthase promoter regulates expression in glial cells.

The expression of inducible nitric oxide synthase (NOS2) in glial cells is inhibited by neurotransmitters such as norepinephrine (NE) which elevate cAMP levels. We examined the molecular basis for this effect using a 2.2-kb fragment of the rat NOS2 promoter transfected into rat C6 glioma cells. Promoter activation (up to six-fold) by lipopolysaccharide (LPS) and interferon-gamma (IFNgamma) was reduced by NE, which alone had no effect. However, a promoter construct extending to bp -130 and containing the proximal nuclear factor-kappa B (NF-kappaB) binding site was minimally activated by LPS and cytokines, but activated up to three-fold by NE. Deletion analysis identified a 27-bp region (bp -187 to -160) as critical for mediating this suppressive effect. This region also enhanced promoter activation by LPS and cytokines, and prevented activation by NE alone. Gel shift analysis revealed constitutive binding to this region, and induction by NE of additional complexes which could be blocked by an antibody against CREB. NE also increased levels of the IkappaBalpha protein which could contribute to its suppressive effects. These results identify a critical role for this 27-bp region in regulation of NOS2 promoter activation and suppression by cAMP.

Lung uptake of antibodies to endothelial antigens: key determinants of vascular immunotargeting.

Vascular immunotargeting is a mean for a site-selective delivery of drugs and genes to endothelium. In this study, we compared recognition of pulmonary and systemic vessels in rats by candidate carrier monoclonal antibodies (MAbs) to endothelial antigens platelet endothelial cell adhesion molecule (PECAM)-1 (CD31), intercellular adhesion molecule (ICAM)-1 (CD54), Thy-1.1 (CD90.1), angiotensin-converting enzyme (ACE; CD143), and OX-43. Tissue immunostaining showed that endothelial cells were Thy-1.1 positive in capillaries but negative in large vessels. In the lung, anti-ACE MAb provided a positive staining in 100% capillaries vs. 5-20% capillaries in other organs. Other MAbs did not discriminate between pulmonary and systemic vessels. We determined tissue uptake after infusion of 1 microg of (125)I-labeled MAbs in isolated perfused lungs (IPL) or intravenously in intact rats. Uptake in IPL attained 46% of the injected dose (ID) of anti-Thy-1.1 and 20-25% ID of anti-ACE, anti-ICAM-1, and anti-OX-43 (vs. 0.5% ID of control IgG). However, after systemic injection at this dose, only anti-ACE MAb 9B9 displayed selective pulmonary uptake (16 vs. 1% ID/g in other organs). Anti-OX-43 displayed low pulmonary (0.5% ID/g) but significant splenic and cardiac uptake (7 and 2% ID/g). Anti-Thy-1.1 and anti-ICAM-1 displayed moderate pulmonary (4 and 6% ID/g, respectively) and high splenic and hepatic uptake (e.g., 18% ID/g of anti-Thy-1.1 in spleen). The lung-to-blood ratio was 5, 10, and 15 for anti-Thy-1.1, anti-ACE, and anti-ICAM-1, respectively. PECAM antibodies displayed low pulmonary uptake in perfusion (2% ID) and in vivo (3-4% ID/g). However, conjugation with streptavidin (SA) markedly augmented pulmonary uptake of anti-PECAM in perfusion (10-54% ID, depending on an antibody clone) and in vivo (up to 15% ID/g). Therefore, ACE-, Thy-1.1-, ICAM-1-, and SA-conjugated PECAM MAbs are candidate carriers for pulmonary targeting. ACE MAb offers a high selectivity of pulmonary targeting in vivo, likely because of a high content of ACE-positive capillaries in the lungs.

Local anesthetics potentiate nitric oxide synthase type 2 expression in rat glial cells.

Expression of the calcium-independent nitric oxide synthase (NOS2) contributes to damage in neurologic disease and trauma. The effects of local anesthetics on NOS2 expression have not been examined. The authors tested the effects of four local anesthetics on the expression of NOS2 in immunostimulated rat C6 glioma cells. Incubation with local anesthetics alone did not induce nitrite accumulation; however, the nitrite production induced by stimulation with bacterial endotoxin lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) was increased in a dose-dependent manner by bupivacaine (maximal 3-fold at 360 microM), tetracaine (maximal 7-fold at 360 microM), and lidocaine at higher doses (5-fold increase at 3.3 mM). Significant increases in nitrite production were observed in concentrations of bupivacaine or tetracaine as low as 120 microM, which correspond to 30 microg/mL (.003% weight/volume). In contrast, ropivacaine had little effect on nitrite production (160% of control values) and only at the highest concentration (3.3 mM, corresponding to 890 microg/mL or 0.089% w/v) tested. Increased nitrite production was not caused by cytotoxic effects of the drugs used, as assessed by release of intracellular lactate dehydrogenase. Increased nitrite production was accompanied by increased NOS2 catalytic activity, steady state mRNA levels, and promoter activation. These results demonstrate that submillimolar doses of two commonly used local anesthetics can increase glial NOS2 expression.

A targetable, injectable adenoviral vector for selective gene delivery to pulmonary endothelium in vivo.

Adenoviral (Ad) vectors are promising gene therapy vehicles due to their in vivo stability and efficiency, but their potential utility is compromised by their restricted tropism. Targeting strategies have been devised to improve the efficacy of these agents, but specific targeting following in vivo systemic administration of vector has not previously been demonstrated. The distinct aim of the current study was to determine whether an Ad-targeting strategy could maintain fidelity upon systemic vascular administration. We used a bispecific antibody to target Ad infection specifically to angiotensin-converting enzyme (ACE), which is preferentially expressed on pulmonary capillary endothelium and which may thus enable gene therapy for pulmonary vascular disease. Cell-specific gene delivery to ACE-expressing cells was first confirmed in vitro. Administration of retargeted vector complex via tail vein injection into rats resulted in at least a 20-fold increase in both Ad DNA localization and luciferase transgene expression in the lungs, compared to the untargeted vector. Furthermore, targeting led to reduced transgene expression in nontarget organs, especially the liver, where the reduction was over 80%. Immunohistochemical and immunoelectron microscopy analysis confirmed that the pulmonary transgene expression was specifically localized to endothelial cells. Enhancement of transgene expression in the lungs as a result of the ACE-targeting strategy was also confirmed using a new noninvasive imaging technique. This study shows that a retargeting approach can indeed specifically modify the gene delivery properties of an Ad vector given systemically and thus has encouraging implications for the further development of targetable, injectable Ad vectors.

The heat shock response inhibits NF-kappaB activation, nitric oxide synthase type 2 expression, and macrophage/microglial activation in brain.

The heat shock response (HSR) provides protection against stress-induced damage, and also prevents initiation of inflammatory gene expression via inhibition of NFkappaB activation. This article describes experiments demonstrating that the HSR prevents induction of nitric oxide synthase type 2 (NOS2) in rat brain. Twenty four hours after intrastriatal injection of lipopolysaccharide (LPS), IL-1beta, and IFN-gamma, NOS2 immunoreactive cells were detected in striatum, corpus callosum, and to a lesser extent in cortex. Induction of a HSR by whole body warming to 41 degrees C for 20 minutes, done 1 day before LPS plus cytokine injection, reduced the number of NOS2-positive staining cells to background levels. Staining for EDI antigen revealed that the HSR also suppressed microglial/brain macrophage activation in the same areas. Striatal injection of LPS and cytokines induced the rapid activation of NFkappaB, and this activation was prevented by prior HS, which also increased brain IkappaB-alpha expression. These results suggest that establishment of a HSR can reduce inflammatory gene expression in brain, mediated by inhibition of NFkappaB activation, and may therefore offer a novel approach to treatment and prevention of neurological disease and trauma.

Inhibitory and stimulatory effects of lactacystin on expression of nitric oxide synthase type 2 in brain glial cells. The role of Ikappa B-beta.

Expression of inflammatory nitric oxide synthase (NOS2) is mediated by transcription factor NFkappaB. By using the specific proteasome inhibitor lactacystin to examine IkappaB degradation, we observed a paradoxical increase in lipopolysaccharide- and cytokine-dependent NOS2 expression at low concentrations or when lactacystin was added subsequent to cytokines. Lactacystin reduced the initial accumulation of NOS2 mRNA but reduced its subsequent decrease. Lactacystin increased NOS2 promoter activation after 24 h, but not after 4 h, and similarly prevented initial NFkappaB activation and at later times caused NFkappaB reactivation. Lactacystin reduced initial degradation of IkappaB-alpha and IkappaB-beta, however, at later times selectively increased IkappaB-beta, which was predominantly non-phosphorylated. Expression of full-length rat IkappaB-beta, but not a carboxyl-terminal truncated form, inhibited NOS2 induction and potentiation by lactacystin. Lactacystin increased IkappaB-beta expression in the absence of NOS2 inducers, as well as expression of heat shock protein 70, and the heat shock response due to hyperthermia increased IkappaB-beta expression. These results suggest that IkappaB-beta contributes to persistent NFkappaB activation and NOS2 expression in glial cells, that IkappaB-beta is a stress protein inducible by hyperthermia or proteasome inhibitors, and that delayed addition of proteasome inhibitors can have stimulatory rather than inhibitory actions.

Purification of radiolabeled monoclonal antibodies to angiotensin-converting enzyme significantly improves specificity and efficacy of its targeting into the lung.

The aim of this study was to improve the labeling/purification procedures for monoclonal antibody (MoAb) to angiotensin-converting enzyme (ACE). MoAb 9B9 was very stable upon iodination at a wide range of iodogen concentrations and incubation times, and was also very stable upon storage, indicating the high technological potential of this MoAb. Radiolabeled MoAb 9B9 was purified by (i) adsorption chromatography on cellulose, (ii) HPLC (gel filtration) and (iii) affinity chromatography on ACE-Sepharose. The best result was obtained with cellulose: specificity of MoAb 9B9 accumulation in the lung increased 2-fold. We conclude that the phenomenon of specific lung accumulation of MoAb 9B9 may serve as an ideal (convenient, cheap and technological) assay system for evaluation of monoclonal antibody modification and labeling.